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Team:DTU-Denmark/Notebook/2 August 2013
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*Gel purification of Cyc and Nir | *Gel purification of Cyc and Nir | ||
*USER reaction | *USER reaction | ||
| + | *PCR of Nir, His-Tag Sec, His-Tag TAT | ||
| + | *Miniprep re-purification with columns | ||
| + | |||
==Who was in the lab== | ==Who was in the lab== | ||
<hr/> | <hr/> | ||
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==Procedure== | ==Procedure== | ||
<hr/> | <hr/> | ||
| + | * Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit. | ||
| + | * USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence. | ||
| + | * PCR according to standard PCR protocol. | ||
| + | Samples names: | ||
| + | * 1,2 - Nir 48 (primers 48a, 48b) on template of Nir (too long fragment amplified at the beginning of work with Nir PCR) | ||
| + | * 3,4 - Sec His-Tag, primers 19a, 19b; template: Sec 2 | ||
| + | * 5,6 - Tat His-Tag, primers 20a, 20b; template: TAT 2 1 | ||
| + | * 7,8 - Tat His-Tag, primers 20a, 20b; template: TAT 3 2 | ||
| + | * 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a | ||
| + | * 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag | ||
==Results== | ==Results== | ||
Revision as of 15:20, 2 August 2013
2 August 2013
Contents |
lab 208
Main purpose
- Gel purification of Cyc and Nir
- USER reaction
- PCR of Nir, His-Tag Sec, His-Tag TAT
- Miniprep re-purification with columns
Who was in the lab
Gosia, Kristian, Natalia, Julia
Procedure
- Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.
- USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.
- PCR according to standard PCR protocol.
Samples names:
- 1,2 - Nir 48 (primers 48a, 48b) on template of Nir (too long fragment amplified at the beginning of work with Nir PCR)
- 3,4 - Sec His-Tag, primers 19a, 19b; template: Sec 2
- 5,6 - Tat His-Tag, primers 20a, 20b; template: TAT 2 1
- 7,8 - Tat His-Tag, primers 20a, 20b; template: TAT 3 2
- 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
- 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag
Results
Conclusion
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